The CGP1 assay measures the decrease in NADPH absorbance at 340 nm during the oxidation of NADPH to NADP+. This is a kinetic assay and the A340/MIN means absorbance per minute. Please see step 4 on page 4 of the technical bulletin for the kinetic program instructions. The number of readings being 6 for an interval time of 10 seconds which means data is for a minute. NADP+ does not have any absorbance at 340 nm and NADPH does. Thus, the absorbance will decrease in a sample with glutathione peroxide as all substrates, cofactors, enzymes are present for this conversion to occur.
CGP1
Glutathione Peroxidase Cellular Activity Assay Kit
Sufficient for 100 colorimetric tests
Sinónimos:
Cellular glutathione peroxidase, c-GPx
About This Item
Productos recomendados
Nivel de calidad
uso
sufficient for 100 tests
caducidad
Kit is stable for 24 months when unopened.
método de detección
colorimetric
Condiciones de envío
wet ice
temp. de almacenamiento
−20°C
Descripción general
Idoneidad
Principio
Otras notas
Solo componentes del kit
- tert-Butyl hydroperoxide 1 mL
Producto relacionado
Palabra de señalización
Danger
Frases de peligro
Clasificaciones de peligro
Acute Tox. 2 Inhalation - Acute Tox. 3 Dermal - Acute Tox. 4 Oral - Aquatic Chronic 2 - Eye Dam. 1 - Flam. Liq. 3 - Muta. 2 - Org. Perox. F - Skin Corr. 1C - Skin Sens. 1 - STOT SE 3
Órganos de actuación
Respiratory system
Código de clase de almacenamiento
5.2 - Organic peroxides and self-reacting hazardous materials
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Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.
Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.
Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.
Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.
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"Does A340/MIN refer to the absorbance value per minute, or is it the rate of decrease per minute?"
1 answer-
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Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be used with Serum samples?
1 answer-
The Glutathione Peroxidase Cellular Activity Assay Kit (Product number: CGP1) is designed to measure Glutathione Peroxidase activity in cell lysates and tissue homogenates. This kit has not been validated for use with serum samples.
Please see product MAK437, Glutathione Peroxidase Assay Kit, which can be used for serum samples:
https://www.sigmaaldrich.com/product/sigma/mak437Helpful?
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Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be run in a 96 well plate?
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Yes, but you would need to have a plate reader that has a kinetic function as this is a kinetic assay.
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What dilution should I use for my sample when using Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?
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In general, when human erythrocytes were assayed for the enzyme, the sample was diluted 10-fold (15 mg peroxidase per mL) and a 20 μl sample gave a decrease of 0.032 OD340 per minute. Too much enzyme will underestimate the apparent activity of the sample. When rabbit reticulocytes were assayed at a 10-fold dilution (10 mg peroxidase per mL), 20 μl gave a decrease of 0.065 OD340 per minute.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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What type of readings do I get with Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?
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There is a decrease in absorbance during the assay. The OD values decrease with the increase in reaction time. The slope of the reduction in OD (delta OD per minute) is the value used to determine the enzyme units.
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How should I prepare my sample for analysis with Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?
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The cells must be lysed before assaying with this kit. The best way to perform lysis is to incubate the cells with hypotonic buffer (at least 2 volumes and better with 3-4 volumes, depending on the expected level of peroxidase in these cells). Let them swell and homogenize with a Dounce homogenizer. Centrifuge the homogenate at 10,000 × g and use the supernatant for the assay. If a detergent is used for lysis, only use a peroxide-free detergent and follow the instructions in the Bulletin: "Nonionic detergents such as TWEEN and TRITON X-100 that contain high levels of endogenous peroxides will raise the apparent activity. If these detergents are vital to the extraction of the proteins of interest, a low peroxide detergent should be used such as Product Codes X-100-PC (TRITON X-100), P 6585 (TWEEN 20), or P 8192 (TWEEN 80)".
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Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be used with tissue samples?
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We have not tested the CGP1 assay with tissue samples. However, we suggest that the customer perform a small-scale pilot experiment using our CelLytic-MT lysis buffer (Product No. C3228). The buffer enables efficient extraction of tissue proteins. It does not contain any of the interfering agents that are listed on the kit bulletin.
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